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Sequence-dependent pausing of single lambda exonuclease molecules
Sequence-dependent pausing of single lambda exonuclease molecules.
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Lambda exonuclease processively degrades one strand of duplex DNA, moving 5'-to-3' in an ATP-independent fashion. When examined at the single-molecule level, the speeds of digestion were nearly constant at 4 nanometers per second (12 nucleotides per second), interspersed with pauses of variable duration. Long pauses, occurring at stereotypical locations, were strand-specific and sequence-dependent. Pause duration and probability varied widely. The strongest pause, GGCGAT TCT, was identified by gel electrophoresis. Correlating single-molecule dwell positions with sequence independently identified the motif GGCGA. This sequence is found in the left lambda cohesive end, where exonuclease inhibition may contribute to the reduced recombination efficiency at that end.
Perkins TT, Dalal RV, Mitsis PG, Block SM
Science (New York, N.Y.)
2003-09-26 00:00
301
5641
1914-8
Bacteriophage lambda,Base Pairing,Base Sequence,Binding Sites,Consensus Sequence,DNA,Electrophoresis, Polyacrylamide Gel,Exodeoxyribonucleases,Hydrogen Bonding,Kinetics,Models, Chemical,Oligodeoxyribonucleotides,Polymerase Chain Reaction,Probability,Stochastic Processes,Time Factors,Viral Proteins,Oligodeoxyribonucleotides,Viral Proteins,DNA,Exodeoxyribonucleases,lambda exonuclease
Department of Biological Sciences, Stanford University, Stanford, CA 94305, USA tperkinsjilacoloradoedu
Science
NIGMS GM 57035, NHGRI HG 011821-01, NIGMS R01 GM057035-08
1095-9203
10.1126/science.1088047
1088047
1005
True
12947034
